PCR methods are known from the prior art. The patent specification U.S. Pat. No. 4,683,202 B1 discloses a method, with which at least one specific nucleic acid sequence contained in a nucleic acid or a mixture of nucleic acids can be amplified, wherein each nucleic acid consists of two separate, complementary strands, of equal or unequal length. The method comprises: (a) treating the strands with two primers for each different specific sequence being amplified under such conditions that, for each different sequence being amplified, an extension product for each primer is synthesized, which is complementary to the respective nucleic acid strand. Said primers are selected so that they are substantially complementary to different strands of each specific sequence, so that the extension product that is synthesized from a primer can be used, if separated from its complement, as a template for the synthesis of the extension product of the other primer; (b) separating the primer extension products from the templates, on which they were synthesized so that single-stranded molecules are produced; (c) treating the single-stranded molecules from step (b) with the primers from step (a) under such conditions that a primer extension product is synthesized, wherein each of the single strands of step (b) is used as a template. The steps can be carried out one after the other or simultaneously. In addition the steps (b) and (c) can be repeated until the desired degree of sequence amplification is achieved.
In the international application laid open for public inspection WO 2007/143034 A1, methods are disclosed that are to be suitable for performing a PCR method. The methods may include the use of an optical radiation source for heating in a PCR method. The methods may also include the use of surface plasmon resonance or fluorescence resonance energy transfer for monitoring a PCR method in real-time. The methods may further include the immobilization of a template, primer or a polymerase on a surface such as gold or another surface that is active in relation to the surface plasmon resonance.
The patent application US 2002/0061588 A1 discloses methods for making nucleic acids locally and directly responsive to an external signal. The signal acts only on one or a plurality of specific localized portions of the nucleic acid. According to the invention the signal can change the properties of a specific nucleic acid and thereby also change its function. Accordingly the invention provides methods for regulating the structure and functioning of a nucleic acid in a biological sample without influencing other constituent parts of the sample. In one embodiment a modulator transfers heat to a nucleic acid or a part of a nucleic acid, which results e.g. in intermolecular or intramolecular bonds being destabilized, and the structure and stability of the nucleic acid changing. Preferred modulators include metal nanoparticles, semiconductor nanoparticles, magnetic nanoparticles, oxide nanoparticles and chromophores. It is also proposed to use these methods in association with a PCR method. It is proposed in particular to control a PCR reaction with a modulator.
The patent application DE 10 2012 201 475 A1 relates to a method for the amplification of nucleic acids. In this method, electromagnetically excited nanoparticles in a reaction volume transfer heat to their environment through excitation. If the heat input is below a critical duration, which depends on the average particle distance in the solution and thus the concentration of the nanoparticles, a very rapid denaturing can be achieved, wherein the duration of the excitation of the nanoparticles is very much shorter than the cycle duration.
The patent DE 10 2013 215 166 B3 (publication date of the grant of patent: 30 Oct. 2014) of the inventors of this patent application contains a method for super-amplification, wherein the shortening of the cycle duration leads to a low yield per cycle, but which is more than compensated by the possibility of being able to perform more cycles per time unit.
The patent application US 2003/0143604 A1 relates to the use of nanoparticle detection probes for monitoring amplification reactions, in particular PCR. The patent application deals primarily with the use of nanoparticle-oligonucleotide conjugates which are treated with a protective reagent such as bovine serum albumen, in order to detect a target polynucleotide quantitatively and qualitatively. The patent application discloses a nucleic acid amplification and detection using gold nanoparticle primers. In a first step the nucleic acid target is denatured in the presence of the gold nanoparticles, to which primers are attached. In a second step the gold nanoparticles hybridize with the primers attached thereto to the nucleic acid target and a copy of the complementary DNA sequence is produced based on the nucleic acid primers which are attached to the nanoparticles. The first and second steps are repeated and the optical signal which is produced through the binding of complementary nanoparticle probes that have been amplified is measured.
The patent specification EP 1 842 924 B1 discloses a method for determining an initial concentration of nucleic acids using nucleic acid real-time amplification data, wherein a measured fluorescence, due to the amplification, passes through a function dependent on the number of cycles passed through.